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Anti goat

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Anti-goat is a laboratory reagent used to detect the presence of goat-derived proteins or antibodies in a sample. It functions by binding to and labeling these targets, allowing for their identification and quantification through various analytical techniques.

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Lab products found in correlation

Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Nitrocellulose membrane, by GE Healthcare (1 mentions) Ne per nuclear and cytoplasmic extraction reagent kit, by Thermo Fisher Scientific (1 mentions) Bis tris plus gel, by Thermo Fisher Scientific (1 mentions) P mtors2448, by Cell Signaling Technology (1 mentions) Lds sample buffer and reducing agent, by Thermo Fisher Scientific (1 mentions) Mtor sc 1549, by Santa Cruz Biotechnology (1 mentions) P pras40 t246, by Cell Signaling Technology (1 mentions) Odyssey infrared detection instrument, by LI COR (1 mentions) Erk 4695, by Cell Signaling Technology (1 mentions) Pakt s473, by Cell Signaling Technology (1 mentions) Ab55391, by Abcam (1 mentions) Sc 20687, by Santa Cruz Biotechnology (1 mentions) Imaging system, by LI COR (1 mentions) Anti mouse irdye conjugated secondary antibodies, by LI COR (1 mentions) Sc 1616, by Santa Cruz Biotechnology (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions)

2 protocols using anti goat

1

Subcellular Fractionation and Western Blot Analysis

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Following treatment, MCF7, MCF7-TamR, ZR75-1, T47D, MDA-MB-231, and MDA-MB-436 cells were harvested and cytoplasmic and nuclear fractionation was performed using NE-PER® Nuclear and Cytoplasmic Extraction Reagent kit (ThermoFisher Scientific, Hampton, NH) according to the manufacturer’s instructions. Samples were subsequently denatured using LDS Sample buffer and Reducing agent (Invitrogen, Carlsbad, CA) at 70 °C for 10 min. Samples were resolved using Bis–Tris Plus gels (Invitrogen, Carlsbad, CA) and transferred onto nitrocellulose membrane (GE Healthcare, Port Washington, NY). Membranes were probed with the following primary antibodies: ERα (sc8005), raptor (sc81537), and mTOR (sc-1549) (Santa Cruz Biotechnology, Dallas, TX), p-ERK (M8159) Sigma-Aldrich (St Louis, MO), PARP (Abcam, Cambridge, UK), p-mTOR-S2448 (5536S), p-Akt-S473 (4060L), Akt (4691X), p-PRAS40-T246 (13175P), p-S6K-T389 (9206S), 4EBP1 (9644S), p-4EBP1-S65 (9454S), and ERK (4695) (Cell Signaling Technologies (Danvers, MA). Signal detection and quantification were accomplished using IRDye-conjugated anti-rabbit (LI-COR, 827-08365, Lincoln, NE), anti-mouse (LI-COR, 926-68070, Lincoln, NE), or anti-goat (LI-COR, 926-68074, Lincoln, NE) secondary antibodies using Odyssey infrared detection instrument (LI-COR, Lincoln, NE). All immunoblots were performed at least thrice to ensure reproducibility.
Bostner J., Alayev A., Berman A.Y., Fornander T., Nordenskjöld B., Holz M.K, & Stål O. (2017). Raptor localization predicts prognosis and tamoxifen response in estrogen receptor-positive breast cancer. Breast Cancer Research and Treatment, 168(1), 17-27.
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2

Western Blot Analysis of Proteins

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Total proteins from tissues were extracted by homogenizing in RIPA buffer premixed with protease inhibitor cocktail (Sigma, St. Louis, MO). Proteins concentrations were determined using a BCA protein assay kit (Thermo Scientific, Rockford, IL). Total protein (50 μg) was separated on 12% mini PROTEAN polyacrylamide gels and then transferred to polyvinylidenefluoride (PVDF) (Life Technologies Carlsbad, CA) using iBlot gel transfer system. Membrane was blocked using Odyssey blocking buffer for 1 h at room temperature. Membranes were incubated overnight with rabbit polyclonal to Gli-1 (SC-20687), rabbit polyclonal to Shh (SC-h160), goat polyclonal to β-actin (SC-1616) (1:1000) (Santa Cruz Biotech., Dallas, TX), and mouse monoclonal antibodies against KRAS (ab-55391) (1:1000) (Abcam, Cambridge, MA). After washing with TBST buffer, the membrane was further incubated with their corresponding antirabbit, antigoat, and antimouse IR dye conjugated secondary antibodies (1:10000) (LI-COR Biosciences, Lincoln, NE) for 60 min and visualized using LI-COR imaging system. Expression levels of desired protein were normalized against β-actin (SC-1616) protein expression levels.
Kumar V., Mondal G., Slavik P., Rachagani S., Batra S.K, & Mahato R.I. (2015). Codelivery of Small Molecule Hedgehog Inhibitor and miRNA for Treating Pancreatic Cancer. Molecular pharmaceutics, 12(4), 1289-1298.
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